Thesis (M.S.)--Oregon State University, 2007. / Printout. Includes bibliographical references (leaves 47-54). Also available on the World Wide Web.

In common bean (Phaseolus vulgaris), the main seed storage pests are the bruchid beetles. Damage done to the seed by the larvae has a large impact on seed quality and yield. Arcelin (ARC), phytohaemagglutinin (PHA), and α-amylase inhibitor (α-AI) are linked seed storage proteins that form the APA locus on chromosome Pv04 and are associated with resistance. A major breeding objective is to introduce bruchid resistance into common bean from a resistant tepary genotype, G40199, by introgressing the resistant APA locus into susceptible common bean backgrounds. Here we developed a molecular marker that tracks the introgression. A set of PCR primers to the α-amylase inhibitor locus amplified a DNA fragment that showed a 45 base pair insertion in the middle of a lectin Leg_b domain. This enhanced locus characterization and insertion/deletion marker may preclude the need for bruchid resistance screening early in the breeding. / United States. Agency for International Development / United States. Global Hunger and Food Security Initiative (Cooperative Agreement No. EDH-A-00-07-00005-00)

Root rot diseases of bean (Phaseolus vulgaris L.) are a problem wherever they are grown, and are a major constraint to dry edible and snap bean production. Root rot is a primary yield limitation of snap bean production in the US, especially within the top three snap bean producing states of Wisconsin, Oregon and New York. Bean root rot pathogens will be present by the end of the first season even when starting with clean ground. The decline in yield can be relatively slow, so growers might not notice or appreciate the hidden yield cost associated with root rot disease. Traditional methods for disease control such as fungicides, crop rotations, cover crops, seedbed preparations have been proven ineffective (either physically ineffective or economically unviable) against root rot. Therefore, genetic resistance is needed. In order to address the need for genetic resistance to root rot in snap beans, the highly root rot resistant line RR6950, a small seeded black indeterminate type IIIA accession of unknown origin, was crossed with OSU5446, a highly root rot susceptible determinate type I blue lake four-sieve breeding line to produce the RR138 recombinant inbred mapping population. In this study we evaluated the RR138 RI population in the F₆ generation for resistance to Fusarium solani root rot in Oregon and Aphanomyces euteiches root rot in Wisconsin. We also evaluated this population for morphological traits and root structural traits including pod height, pod width, pod length, pod wall thickness, strings, seed color, flower color, tap and basal root diameter, and root angle measurements. The RR138 population was also genotyped on the 10K BeanCAP Illumina Beadchip. The Single Nucleotide Polymorphism (SNP) data was used to assemble a high-density linkage map and Quantitative Trait Loci (QTL) for phenotypic data were evaluated. The linkage map produced from this study contained 1,689 SNPs across 1,196cM. The map was populated with 1 SNP for every 1.4cM, spanning across 11 linkage groups. Three QTL associated with A. euteiches root rot resistance were consistently expressed in 2011 and 2012 trials. A. euteiches QTL were found on Pv02, Pv04, and Pv06 and accounted for 7-17% of total genetic variation. Two QTL associated with F. solani were found in 2011 trial on Pv03 and Pv07, account for 9 and 22% of total genetic variation, respectively. We also found several QTL for morphological traits and root structural traits including QTL for pod fiber and pod height on Pv04, pod length on Pv01, strings on Pv01, taproot diameter on Pv05, and shallow basal root angle on Pv05, accounting for 21, 26, 12, 20, 11, and 19% of total genetic variation, respectively. QTL discovered from Oregon data for F. solani resistance did not cluster with QTL for A. euteiches root rot resistance. "SNP0928_7", was highly associated with F. solani resistance on Pv07 and "SNP0508_2", was highly associated with A. euteiches on Pv02. QTL and markers associated with QTL from this study will be of value to snap bean breeders developing root rot resistant lines with processing traits, and provide more information about targeting the mechanism of resistance. / Graduation date: 2013

Thesis (M. Sc.)(Microbiology)--University of Pretoria, 2005. / Includes bibliographical references. Available on the Internet via the World Wide Web.

Common bean is a nutritionally and economically important food crop and a major source of dietary protein in many developing countries throughout the world. Seed coat colour and size in this crop are the main factors determining its marketability in different parts of the world. Flavonoid compounds that are responsible for seed coat colour in beans have been shown to have anti-oxidant, anti-proliferative, anti-tumor, anti-inflammatory, and pro-apoptotic activities. They also may enhance the resistance of beans to pest and disease. A better understanding of the relationships between seed coat colour and flavonoid metabolism in the seed coat may help breeders to select for more nutritionally-beneficial bean varieties. The objective of this research was to test the hypothesis that the genes determining colour in beans are structural and regulatory genes of the phenylpropanoid pathway. The map positions of phenylpropanoid genes were determined in two recombinant inbred populations. Segregation patterns of 18 phenylpropanoid pathway genes in the BAT93 × Jalo EEP 558 RIL population and five phenylpropanoid pathway genes in OAC Rex × SVM Taylor were used to place them on the linkage maps for these populations. Five out of 18 genes were mapped within 2-17 cM of colour gene loci in the BAT93 × Jalo EEP 558 RIL population. The sequences of central genes of the phenylpropanoid pathway were determined by sequencing 6 BAC clones selected with probes for two PAL genes, two CHS genes, DFR, and Myb. The functional annotations of the BAC clones were determined and the similarities between bean phenylpropanoid genes and their corresponding orthologs in other plant species were investigated. A recently developed approach of whole genome sequence comparison was utilized to compare the microsynteny of the sequenced BAC clones with regions of the soybean genome. The physical locations of BAC clones were verified on the bean genome and their counterpart locations on the soybean genome were confirmed. The results agreed with previous studies that indicated that bean genome segments have two homologous segments in soybean and confirmed the high degree of microsynteny that is shared between bean and soybean.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1948. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves [82]-84).

Thesis (M.S.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 64-72). Also available on the World Wide Web.

Made available in DSpace on 2015-03-26T13:42:12Z (GMT). No. of bitstreams: 1 texto completo.pdf: 831305 bytes, checksum: 7de0dccf59e4128a858ec0de24b05804 (MD5) Previous issue date: 2009-08-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Common bean (Phaseolus vulgaris L.) is a crop of great nutritional, economical and social importance. Breeding programs use molecular markers as important auxiliary tools for various types of genetic studies. Different classes of molecular markers have been developed, and among them microsatellites highlight. Microsatellites are DNA simple sequence repeats (SSR) distributed in tandem along the genome, forming highly variable polymorphic sites, enabling their use as molecular markers. SSRs are codominant, multiallelic, and thus can be used in several types of studies, mainly for genetic mapping. Primers flanking microsatellite sequences are commonly developed from genomic libraries, enriched genomic libraries, sequences obtained from databases and, alternatively, from internal simple sequence repeats (ISSR-PCR). Common bean breeding molecular geneticists have developed SSR markers for mapping specific traits of interest. However, a saturated consensus genetic map for common bean has not been established so far that can be used as a reference for the development of specific maps. Therefore, the BIOAGRO/UFV common bean breeding program developed a population of recombinant inbred lines (RILs) which is suggested to be used to integrate, in one single map, all microsatellite markers that have been developed so far. However, to saturate the map, there must be a great number of markers available. The objective of the present study was to develop and validate primers that amplify microsatellites sequences obtained from enriched genomic libraries and from ISSR sequences. In the first case, two enrichedgenomic libraries for microsatellite sequences, that had been developed in a previous study, were used. In the present study, 207 clones were selected from these two genomic libraries. One hundred and ninety six of these clones (94.68%) were sequenced and 184 (88.89%) of them could be analyzed, 133 clones from library 1 and 51 from library 2. Forty eight redundant clones (26.09%) were detected. Clone analysis led to the identification of 66 (49.62%) microsatellite motifs in library 1 and 20 (39.22%) in library 2, and 56 primer pairs were designed. From the 56 primer pairs developed, 34 were characterized andtested in 20 Mesoamerican and Andean genotypes, including AND277 and Rudá. All the primer pairs were able to generate PCR products and six (17.65%) generated polymorphic DNA bands among the tested genotypes. In the ISSR enrichment methodology, 250 clones were seleted with sizes over 400 bp. From these 250 clones, 168 (67.2%) were sequenced and 103 (41.2%) could be analyzed. Thirty redundant clones (29.13%) were detected. Clone analyses led to the identification of 58 microsatellite motifs (56.31%) and 32 primer pairs were developed. Out of these, 10 were characterized and tested in the same genotypes used in the previous methodology. Out of the 10 primer pairs tested, six were identified as codominant markers and the other four as dominant. The codominant markers revealed no polymorphisms among the tested genotypes. Additionally, microsatellite containing sequences obtained from both methodologies were submitted to BLAST analysis against sequences deposited in public databases. Similarity was identified between the SSR sequences and transcribed and non-transcribed regions, from nuclear, mitochondrial and chloroplast genomes, and also with retrotransposon sequences. Both methodologies were effective for selecting, in common bean, sequences that contain microsatellites. The results obtained represent an initial effort to select molecular markers that will be mapped in the future RIL's consensus population, contributing for the construction of a satured genetic map for the species. In addition, these primers can be used in different types of genetic studies which are important for common bean breeding programs that use molecular markers. / O feijão-comum (Phaseolus vulgaris L.) é uma cultura de destacada importância nutricional, econômica e social. Programas de melhoramento genético do feijoeiro têm utilizado marcadores moleculares como importantes ferramentas auxiliares em diversos tipos de estudos genéticos. Diferentes classes de marcadores têm sido desenvolvidas, dentre as quais se destacam os microssatélites. Os microssatélites (SSR) são seqüências simples repetidas de DNA, que se repetem em tandem ao longo do genoma, formando sítios altamente polimórficos, o que possibilita o seu uso como marcas moleculares. Como marcadores, são codominantes, multialélicos, e aplicáveis em diversos tipos de estudos, principalmente no mapeamento genético. Primers que flanqueiam sequências SSR geralmente são desenhados a partir da construção de bibliotecas genômicas, bibliotecas genômicas enriquecidas, sequências depositadas em bancos de dados e, alternativamente, a partir de seqüências internas simples repetidas (ISSR). Geneticistas moleculares têm desenvolvido marcadores SSR com o intuito de mapear genes que codificam determinadas características de interesse. Entretanto, não existe um mapa consenso saturado para o feijão que sirva como referência para auxiliar na construção de mapas específicos. Nesta perspectiva, o Programa de Melhoramento Genético do Feijoeiro do BIOAGRO/UFV desenvolveu uma população de RIL s que poderá ser usada para integrar, em um único mapa, todos os marcadores SSR já desenvolvidos. No entanto, para a saturação do mapa, há necessidade de um grande número de marcadores. O presente trabalho teve o objetivo de desenvolver e validar primers que amplifiquem regiões contendo microssatélites a partir da metodologia da construção de bibliotecas genômicas enriquecidas e a partir de ISSR. Na primeira metodologia, em trabalho anterior, foram construídas duas bibliotecas genômicas enriquecidas para seqüências SSR. No presente trabalho, a partir das bibliotecas genômicas desenvolvidas, foram selecionados 207 clones contendo insertos de tamanho desejado. Destes, foram seqüenciados 196 (94,68%), dos quais 184 (88,89%) puderam ser analisados, sendo 133 clones da biblioteca 1 e 51 da biblioteca 2. Foram detectados 48 (26,09%) clones redundantes. A análise dos clones permitiu identificar 66 (49,62%) motivos SSR na biblioteca 1 e 20 (39,22%) na biblioteca 2, a partir dos quais foram desenhados 56 pares de primers. Destes, 34 tiveram suas condições de amplificação otimizadas e padronizadas e foram testados quanto ao polismorfismo detectado entre 20 genótipos andinos e mesoamericanos, incluindo os genitores AND277 e Rudá. Todos os primers geraram produtos de amplificação e seis (17,65%) amplificaram produtos polimórficos entre os genótipos testados. Em relação à metodologia de enriquecimento por ISSR-PCR foram selecionados 250 clones contendo insertos com tamanho desejado, obtidos a partir da amplificação por ISSRPCR, clonagem dos fragmentos e transformação de células competentes. Dos 250 clones, 168 (67,2%) foram sequenciados e 103 (41,2%) puderam ser analisados. Foram detectados 30 clones redundantes (29,13%). A análise das sequências permitiu identificar 58 motivos microssatélites (56,31%) e foi possível o desenho de 32 pares de primers. Destes, 10 tiveram suas condições de amplificação padronizadas e foram analisados quanto ao polimorfismo detectado entre os mesmos 20 genótipos andinos e mesoamericanos utilizados na metodologia de bibliotecas genômicas enriquecidas. Dos 10 pares de primers testados, seis comportaram-se como marcadores codominantes e quatro como dominantes. Dos codominantes nenhum mostrou-se polimórfico dentre os genótipos testados. Adicionalmente, as sequências contendo motivos microssatélites, obtidas a partir das duas metodologias utilizadas, foram submetidas à busca por similaridade com sequências já caracterizadas em bancos públicos de sequências. Foi identificada similaridade com regiões transcritas e não traduzidas, e com regiões codificadoras de proteínas, a partir do genoma nuclear, mitocondrial e do cloroplasto, e também a partir de sequências advindas de retrotransposons. As duas metodologias utilizadas foram eficazes para a seleção, no feijoeiro, de seqüências contendo microssatélites. Estes resultados representam um primeiro esforço no sentido de selecionar marcadores moleculares que serão futuramente mapeados na população consenso de RILs, além de fornecer marcadores que poderão ser usados nos mais variados tipos de estudos genéticos, contribuindo de fundamental maneira para o aprimoramento dos programas de melhoramento do feijoeiro comum que utilizam marcadores moleculares.

M.Sc. / A common feature of multi gene families is that their members are expressed in different ways in response to environmental and developmental signals. In the present study the expression of a CHS multi gene family in bean (Phase°lus vulgari.), was studied. using a RT-PCR technique that focuses on the 3' divergent regions of the isogenes. Tissue-specific expression in roots, stems, leaves and the flowers of Phaseolus vulgaris, as well as in callus tissue, were investigated. Patterns and levels of gene expression were investigated after treatment with different elicitors as well as light. In most cases time and concentration studies were performed. The four CHS transcripts (CHS4. CHS1. CHS17 and CHS14) showed tissue-specific expression. The four CHS transcripts were differently expressed in the seven organs investigated: and different levels of activity were observed. The highest level of transcript expression for CHS14 and CHS4 could be observed in the roots, whereas relatively low levels were obtained in the leaves. stems and flowers of the green as well as etiolated seedlings. Higher levels of CHS were found in flower buds. High levels of all four transcripts were also found in callus. Elicitor treatment with structurally diverse abiotic agents showed induction of all four CHS mRNA transcripts. Concentration studies revealed high levels of CHS transcript levels. Elicitation with different concentrations of the elicitors: glutathione. mercuric chloride and sodium salicylate showed high levels of the CHS transcripts after exposure of 6 h to the different elicitation agents. The transcript levels increased significantly to levels above those observed in untreated (control) plants. The CHS transcripts showed higher levels of induction after elicitation with mercuric chloride (1 mM) relative to treatment with sodium salicvlate (10 rnM), suggesting differential regulation at the transcriptional level. The expression patterns observed with glutathione were very similarly to those induced by mercuric chloride. The kinetics of induction of all CHS transcripts. except for CHS1 were low at 2 and at 8 h postelicitation and maximal levels of transcript. although transiently induced, could be observed at 4 - 6 h. The use of 4 mM mercuric chloride did not give any induction. most probably because it was a lethal concentration. Etiolated and green bean seedlings, exposed to UV light. showed expression of all four CHS transcripts. In green leaves no significant differences in the induction kinetics between the different chs genes were observed. Three of the transcripts (CHS4. CHSI7. CHS14) accumulated rapidly (within ca. 3h). reaching a maximum after 6 h of irradiation. followed by a decline. CHS4 revealed a 18.2 fold induction. CHSI7 showed a 4.8 fold increase and CHS14 a 4 fold increase after 6 h of illumination in green leaves. In contrast. CHS1 showed a delay ed response which was still observable after 15 h. It was also demonstrated that CHS transcripts accumulated rapidly but transiently. Following illumination of etiolated leaves with white light. except for CHS I. CHS17 and CHS4 showed similar expression levels and patterns. with maximal induction at 1,5 h after white light exposure, whereas maximal induction for CHS14 was at 2 h. At 2.5 Ii the levels for all three transcripts dropped to preinduction levels. It is therefore evident that CHS is a key metabolic control point in the phenylpropanoid pathway leading specifically to isollavonoid biosynthesis. The results strongly suggest that the activation of plant defence genes are regulated in a tissue-specific manner and that induction by different elicitor-active agents. may be regulated by different. But convergent signal transduction regulatory networks.

Experiments were conducted in controlled-environment cabinets to show the effect of temperature and light intensity on the growth and fruiting of snap beans. Leaf weights varied inversely with the temperature, but stem weights and numbers of nodes were not greatly affected by day temperature in the range of 75° to 95°F. Blossoming and pod set were similar at day temperatures of 75° and 85°F but were reduced at 95°F. When day temperature was 95°F, a 60°F night temperature resulted in increased blossoming and pod set compared to 80°F. When pods were harvested at marketable maturity, blossoming in bush beans was cyclic. Plants grown at a light intensity of 1900 foot-candles had a lower fresh and dry weight of leaves, stems, and pods, and fewer blossoms and pods set than plants grown at 2700 and 4000 foot-candles. Field experiments showed that planting dates after May 29 reduced the yield of pods in pole beans. Nitrogen level and row direction did not affect yield of pods in pole beans. Number of pods per plant in pole beans increased directly with the row spacing. Chemical sprays at blossoming caused no increase in yield of pods in pole beans, and only a slight increase in the yield of pods in bush beans. Differences in yields of pods between varieties of bush beans were due differences in the number of pods per plant. / Land and Food Systems, Faculty of / Graduate